dc.contributor.advisor | Gray, Nathanael | |
dc.contributor.advisor | Walker, Suzanne | |
dc.contributor.author | Tan, Zhi Wei | |
dc.date.accessioned | 2020-10-05T12:02:08Z | |
dc.date.created | 2020-03 | |
dc.date.issued | 2020-01-22 | |
dc.date.submitted | 2020 | |
dc.identifier.citation | Tan, Zhi Wei. 2020. Novel OGT Inhibitors Reveal O-GlcNAc Regulates Splicing. Doctoral dissertation, Harvard University, Graduate School of Arts & Sciences. | |
dc.identifier.uri | https://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37365514 | * |
dc.description.abstract | Reversible glycosylation of nuclear and cytoplasmic proteins is an important regulatory mechanism across metazoans. One enzyme, O-linked N-acetylglucosamine transferase (OGT), is responsible for all nucleocytoplasmic protein glycosylation and there is a well-known need for potent, cell-permeable inhibitors to interrogate OGT function. Here we report the structure-based evolution of OGT inhibitors culminating in compounds with low nanomolar inhibitory potency and on-target cellular activity. The structures we report provide insight into how to inhibit glycosyltransferases, a family of enzymes that has been notoriously refractory to inhibitor development.
Intron detention in precursor RNAs serves to regulate expression of a substantial fraction of genes in eukaryotic genomes. How detained intron (DI) splicing is controlled is poorly understood. Here we show that O-GlcNAc, which is thought to integrate signaling pathways as nutrient conditions fluctuate, controls detained intron splicing. Using specific inhibitors of OGT and the enzyme that removes O-GlcNAc (OGA), we first show that O-GlcNAc regulates splicing of the highly conserved detained introns in OGT and OGA to control mRNA abundance in order to buffer O-GlcNAc changes. We show that OGT and OGA represent two distinct paradigms for how DI splicing can control gene expression. We also show that when DI splicing of the O-GlcNAc-cycling genes fails to restore O-GlcNAc homeostasis, there is a global change in detained intron levels. Strikingly, almost all detained introns are spliced more efficiently when O-GlcNAc levels are low, yet other alternative splicing pathways change minimally. Our results demonstrate that O-GlcNAc controls detained intron splicing to tune system-wide gene expression, providing a means to couple nutrient conditions to the cell’s transcriptional regime. | |
dc.description.sponsorship | Chemical Biology | |
dc.format.mimetype | application/pdf | |
dc.language.iso | en | |
dash.license | LAA | |
dc.subject | Small molecule inhibitor development, RNA splicing, proteomics, phosphoproteomics, chemical biology, RNA-Seq, OGT | |
dc.title | Novel OGT Inhibitors Reveal O-GlcNAc Regulates Splicing | |
dc.type | Thesis or Dissertation | |
dash.depositing.author | Tan, Zhi Wei | |
dc.date.available | 2020-10-05T12:02:08Z | |
thesis.degree.date | 2020 | |
thesis.degree.grantor | Graduate School of Arts & Sciences | |
thesis.degree.grantor | Graduate School of Arts & Sciences | |
thesis.degree.level | Doctoral | |
thesis.degree.level | Doctoral | |
thesis.degree.name | Doctor of Philosophy | |
thesis.degree.name | Doctor of Philosophy | |
dc.contributor.committeeMember | Marto, Jarrod | |
dc.contributor.committeeMember | Boutz, Paul | |
dc.type.material | text | |
thesis.degree.department | Chemical Biology | |
thesis.degree.department | Chemical Biology | |
dash.identifier.vireo | | |
dash.author.email | hayzhiwei@gmail.com | |